A method of inducing meiosis in cultured human cells could greatly advance research into human reproduction and lead to new therapies for infertility. Here, we present an in vitro model to induce meiosis starting from male or female human pluripotent stem cells.
Previously published scRNAseq data was used to identify reliable markers for early and late meiotic cells. Then candidate meiosis-promoting factors were screened using scRNA seq. By screening conditions for activating the expression of meiotic genes, we found that DNMT1 inhibition, retinoic acid receptor activation, and overexpression of anti-apoptosis and pro-meiosis factors can rapidly initiate meiosis in male and female hiPSCs. The meiotic cells express genes similar to meiotic oogonia in vivo, including all synaptonemal complex components and machinery for meiotic recombination. These findings establish an accessible system for inducing human meiosis in vitro.
We used Parse scRNAseq in two components of this project. First, we used the WT Mega v2 kit combined with the gene capture kit as a screening tool for identification of pro-meiotic factors. Second, we characterized the gene expression in our cells over the course of our optimized meiosis induction protocol using a Parse WT v3 kit. The ability of Parse technology to store fixed frozen cells prior to large-scale library prep greatly simplified our time course scRNAseq experiment.
The pre-print is available here: https://doi.org/10.1101/2024.05.31.596483
"The ability to store fixed frozen cells greatly simplified our time course experiment."
We're your partners in single cell
Reach out for a quote or for help planning your next experiment.